New phenomenon in early development of sporelings in Gracilaria asiatica Chang et Xia （Gracilariaceae, Rhodophyta）

1 INTRODUCTION The red alga Gracilaria belongs to Gracilari- aceae, Gigartinales, Florideophyceae, and includes over 100 recorded species, wildly distributing in tropical and temperate regions (Oliveira and Plastino, 1994), and is useful for producing val…     

A Comparison of Different Gracilariopsis lemaneiformis （Rhodophyta） Parts in Biochemical Characteristics,Protoplast Formation and Regeneration

Gracilariopsis lemaneiformis is a commercially exploited alga.Its filaceous thallus can be divided into three parts,holdfast,middle segment and tip.The growth and branch forming trend and agar content of these three parts were analyzed,respectively,in this study.The results showed that the tip had the highest growth rate and branched most,although it was the last part with branch forming ability.The holdfast formed branches earliest but slowly.Holdfast had the highest agar content.We also assessed the difference in protoplast formation and regeneration among three parts.The middle segment displayed the shortest enzymolysis time and the highest protoplast yield; whereas the tip had the strongest vitality of protoplasts formation.Juvenile plants were only obtained from the protoplasts generated from the tip.These results suggested that the differentiation and function of G.lemaneiformis was different.     

Cloning and Analysis of Calmodulin Gene from Porphyra yezoensis Ueda （Bangiales, Rhodophyta）

In order to understand the mechanisms of signal transduction and anti-desiccation mechanisms of Porphyra yezoensis, cDNA and its genomic sequence of Calmodulin gene (CaM) was cloned by the technique of polymerase chain reaction (PCR) based on the analysis of P. yezoensis ESTs from dbEST database. The result shows that the full-length cDNA of CaM consists of 603 bps including an ORF encoding for 151 amino acids and a terminate codon UGA, while the length of genomic sequence is 1231 bps including 2 exons and 1 intron. The average GC content of the coding region is 58.77%, while the GC content of the third position of this gene is as high as 82.23%. Four Ca²⁺ binding sites (EF-hand) are found in this gene. The predicted molecular mass of the deduced peptide is 16688.72 Da and the pI is 4.222. By aligning with known CaM genes, the similarity of CaM gene sequence with homologous genes in Chlamydomonas incerta and Chlamydomonas reinhardtii is 72.7% and 72.2% respectively, and the similarity of the deduced amino acid sequence of CaM gene with homologous genes in C. incerta and C. reinhardtii are both 71.5%. This is the first report on CaM from a species of Rhodophyta.     

Biomass and carbon storage of Gracilariopsis lemaneiformis （Rhodophyta） in Zhanshan Bay,Qingdao, China

Marine macroalgae can absorb carbon and play an important role in carbon sequestration. As an important economic macroalga, Gracilariopsis lemaneiformis has the potential to significantly affect carbon absorption and storage in wave-sheltered intertidal reef systems. However, detailed knowledge on seasonal biomass changes and carbon storage of G. lemaneiformis is lacking, especially in many small and scattered ecosystems. Considering the influence of human activities on wild distribution of G. lemaneiformis, the understanding of seasonal dynamics of an economically important species in nature is necessary. In this study, we first investigated seasonal variations in biomass, coverage area, and carbon storage during low tide from August 2011 to July 2012 in Zhanshan Bay, Qingdao, China. Furthermore, we estimated the carbon storage potential of wild G. lemaneiformis using light use efficiency(LUE). The results show that the standing biomass and coverage area changed significantly with season. However, seasonal variations in carbon content and water content were not obvious, with an average content of 35.1% and 83.64%, respectively. Moreover, carbon storage in individual months varied between 0.67 and 47.03 g C/m 2, and the value of carbon storage was the highest in August and June and the lowest in February. In Zhanshan Bay, LUE of G. lemaneiformis was only 0.23%. If it is increased to the theoretical maximum(5%–6%), the carbon storage will have an increase of at least 21 times compared with the current, which suggested that carbon storage of wild G. lemaneiformis had a high enhancement potential. The study will help to assess a potential role of G. lemaneiformis in reducing atmospheric CO2.     

Effects of UV-B irradiation on isoforms of antioxidant enzymes and their activities in red alga Grateloupiafilicina （Rhodophyta）

Macroalgae in a littoral zone are inevitably exposed to UV-B irradiance.We analyzed the effects of UV-B on isoenzyme patterns and activities of superoxide dismutase(SOD),peroxidase(POX),catalase(CAT),and ascorbate peroxidase(APX) of red algae Grateloupia filicina(Lamour.) C.Agardh.The activities of SOD,CAT,and APX changed in response to UV-B in a time- and dose-dependent manner.POX activity increased significantly under all three UV-B treatments.The enzymatic assay showed three distinct bands of SODⅠ(Mn-SOD),SODⅡ(Fe-SOD),and SODⅢ(CuZn-SOD) under a low(Luv) and medium(Muv) dose of UV-B irradiation,while SODⅠ and SODⅢ activities decreased significantly when exposed to a high dose of UV-B irradiation(Huv).The activity of POX isoenzymes increased significantly after exposure to UV-B,which is consistent with the total activity.In addition,a clear decrease in activity of CATIV was detected in response to all the three doses of UV treatments.Some bands of APX isoenzyme were also clearly influenced by UV-B irradiation.Correspondingly,the daily growth rate declined under all the three exposure doses,and was especially significant under Muv and Huv treatments.These data suggest that,although the protection mechanisms of antioxidant defense system are partly inducible by UV-B to prevent the damage,G.filicina has incomplete tolerance to higher UV-B irradiation stress.     

An improved PCR method for direct identification of Porphyra （Bangiales, Rhodophyta） using conchocelis based on a RUBISCO intergenic spacer

An improved method of PCR in which the small segment of conchocelis is amplified directly without DNA extraction was used to amplify a RUBISCO intergenic spacer DNA fragment from nine species of red algal genus Porphyra(Bangiales,Rhodophyta),including Porphyra yezoensis(Jiangsu,China),P.haitanensis(Fujian,China),P.oligospermatangia(Qingdao,China),P.katadai(Qingdao,China),P.tenera(Qingdao,China),P.suborboculata(Fujian,China),P.pseudolinearis(Kogendo,Korea),P.linearis(Devon,England),and P.fallax(Seattle,USA)....     

Ontogenetic development in the morphology and behavior of loach （Misgurnus anguillicaudatus） during early life stages

Loach （Misgurnus anguillicaudatus） is a commercially important fish in China and an ideal aquaculture species. However, culturists experience high larval and juvenile mortality during mass production. To provide insight into ways to improve larviculture techniques, we describe the morphological characteristics and behavior of loach during the larval and early juvenile stages. Yolksae larvae ranged from 2.8 to 4.0 mm body length （BL） between days 0 to 4; preflexion larvae ranged from 3.6 to 5.5 mm BL between days 4 to 6; flexion larvae ranged from 4.8 to 8.1 mm BL between days 5 and 14; and post-flexion larvae ranged from 7. l to 15.7 mm BL between days 11 to 27; the minimum length and age of juveniles was 14.1 mm BL and 23 d, respectively. Loach are demersal from hatch through to the early juvenile stages. A suite of morphological characteristics （e.g., external gill filament and ventral mouth opening） and behavioral traits have developed to adapt to demersal living. We observed positive allometric growth in eye diameter, head length, head height, and pectoral fin length during the early larval stages, reflecting the priorities in the development of the organs essential for survival. Our results provide a basis for developing techniques to improve the survival of larval and juvenile loach during mass production.     

Effects of salinity, light and temperature on growth rates of two species of Gracilaria （Rhodophyta）

Effects of temperature, salinity and light intensity on growth rates of Gracilaria lichenoides and G. tenuistipitata var. liui Zhang et Xia were tested. Eight to ten levels of each factor were first tested separately. The best growth rate was obtained under the conditions of 32°C, 30 and 240 μmol/(m²·s) for G. lichenoides, and 24°C, 20 and 200 μmol/(m²·s) for G. tenuistipitata, respectively. Then a uniform design was used to evaluate the optimal combinations of the three factors. The best conditions for the highest daily specific growth rates (% increase in wet weight) are determined to be 31.30°C, 32.10, and 287.23 μmol/(m²·s) for G. lichenoides (16.26%/d), and 25.38°C, 21.10, and 229.07 μmol/(m²·s) for G. tenuistipitata (14.83%/d), respectively. Supported by the 908 Special Program (908-02-04-07), the National Basic Research Program of China (973 Program, No. 2006CB400608), and K. C. Wong Magna Fund in Ningbo University     

Effect of temperature and photoperiod on maturation of Cephalothrix hongkongiensis （Nemertea： Palaeonemertea）

The limited knowledge about the relationships between environmental factors and the maturation and reproductive seasonality of nemerteans is mostly obtained from field studies, and in many cases is conjectural. Few species have been studied under well-controlled laboratory conditions. In this work, the effects of temperature and photoperiod on gonad development and spawning of Cephalothrix hongkongiensis were observed by rearing worms under different temperatures and photoperiods. Worms reared at 4℃ and 6℃ showed no growth or gonadal development, and all died within 109 days. At 8℃ and 12℃, gonads could be seen from 90 days to the termination of the 360-day experiment, but no spawning was observed. Worms maintained at 16, 24, and 28℃ showed both gonadal development and spawning, and matured faster at higher temperatures. Under a fixed temperature(16℃), photoperiod did not have any significant impact on maturation and spawning. It is likely that temperature plays a major role in regulating the maturation and reproductive cycle of C. hongkongiensis.     

Early Development of Silvetia babingtonii （Fucales,Phaeophyceae）

Silvetia babingtonii is a potentially economic brown alga for sources of food and high-value added utilization.So far,sporeling nursery and field cultivation has not been successful.The lack of knowledge on development and life cycle of this alga hinder the development of techniques for the sporeings and cultivation.In this study,internal structure of oogonium and antherium of S.babingtonii was observed with hematoxylin and eosin staining and through microscope.Meanwhile,early development from zygotes to juvenile sporelings was studied at 20℃ under 60-100 μmol photons m^-2 s^-1.Zygotes germinated and divided into thallus and rhizoid cells.The larger thallus cells further divided and developed into juvenile sporelings; while the smaller rhizoid cells divided and elongated into rhizoid hairs.These findings documented the life cycle ofS.babingtonii and provided fundamental knowledge for sporeling nursery in the near future.     

Formation and early development of tetraspores of Polysiphonia urceolata (Rhodomelaceae, Rhodophyta)

Polysiphonia urceolata is one type of potential commercial red seaweeds used for breeding and cultivation, because of its significant biochemical and biomedical application. However, the information of breeding and seedling incubation for cultivation is limited, especially the early development. In this study, tetrasporohyte and gametophyte of P. urceolata were taken as the study materials in Huiquan Bay, Qingdao, China. The cleaned and sterilized tetrasporophytes and gametophytes were pre-cultured in sterilized seawater, then nurtured at 18℃, 25 μmol photons m-2 s-1 in 12:12 h (light:dark) photoperiod. Continuous observation under microscope showed that the early development consists of bipolar division stage and seedling stage. In the division stage, tetraspores germinate into bipolar sporelings that further differentiate into a colorless rhizoidal portion and a lightly pigmented upright shoot. The lightly pigmented rhizoidal cell develops to a rhizoid and the larger pigmented cell transforms to an erect axis. In the seedling stage, several quasi-protuberances appear on the erect axis and form juvenile seedlings. The results demonstrate the culture of P. urceolata from tetraspores under laboratory conditions.     

Effect of temperature and irradiance on the growth and reproduction of Enteromorpha prolifera J. Ag. （Chlorophycophyta,Chlorophyceae）

Effect of temperature and irradiance on growth and reproduction of Enteromorpha prolifera that bloomed offshore along the Qingdao coast in summer 2008, was studied. It was showed that E. prolifera propagated mainly asexually with specific growth rate （SGR） of 10.47 at 25℃/40 μmol m^-2s^-1. Under this condition, gametes with two flagellate formed and released in 5 days. At the beginning of the development, the unicell gamete divided into two cells with heteropolarity, and then the apical cell developed into thalli primordial cells, whereas the basal cell developed into rhizoid primordial cells. In 8-day culture, the monoplast gamete developed into juvenile germling of 240 μm in length. Unreleased gametes can develop directly within the alga body. E. prolifera could either reproduce through lateral branching or fragmenting except apomixis revealed by Microscopic observation. On aged tissue of E. prolifera, although the degraded pigments partially remained in faded algal filaments, numerous vegetative cells could still divide actively in the algal tissues.     

Induction and cultivation of cloned filaments of Polysiphonia urceolata (Rhodomelaceae, Rhodophyta)

A filamentous clone of Polysiphonia urceolata was regenerated from segments cut from the fronds of gametophytes. Unlike wild thalli with short virgate branchlets, the clone was filamentous with few branches. Many transparent trichoblasts arose from pericentral cells during the induction culture, but these were seldom observed during normal growth. The trichoblasts were uniseriate, often colorless, and formed lobed rhizoids rapidly when they came into contact with solid substrates. In addition to morphological characteristics, the photosynthetic properties and growth conditions of the clone differed from those of the mother plant. Cross-gradient light and temperature culture experiments revealed that the most favorable conditions for culture of the filamentous clone were 22°C and 95-120 μE/(m2-s) light intensity. The photosynthetic light saturation value for filaments was approx. 100 μE/(m2-s), which is far lower than that of wild thalli. These results could be used to develop techniques for mass cultures of P. urceolata in photobioreactors for production of seed stock or bioactive products.     

Inheritance mode of microsatellite loci and their use for kinship analysis in the Pacific oyster （Crassostrea gigas）

Five full-sib families of the Pacific oyster （Crassostrea gigas） larvae were used to study the mode of inheritance at eight microsatellite loci, and the feasibility of these markers for kinship estimate was also examined. All eight microsatellite loci were compatible with Mendelian inheritance. Neither evidence of sex-linked barriers to transmission nor evidence of major barriers to fertilization between gametes from the parents was shown. Three of the eight loci showed the presence of null alleles in four families, demonstrating the need to conduct comprehensive species-specific inheritance studies for microsatellite loci used in population genetic studies. Although the null allele heterozygotes were considered as homozygotes in the calculation of genetic distance, offspring from five full-sib families were unambiguously discriminated in the neighbor-joining dendrogram. This result indicates that the microsatellite markers may be capable of discriminating between related and unrelated oyster larvae in the absence of pedigree information, and is applicable to the investigation of the effective number of parents contributing to the hatchery population of the Pacific oyster.     

Genetic improvement on Chinese shrimp （Fenneropenaeus chinensis）： growth and viability performance in F1 hybrids of different populations

Fenneropenaeus chinensis distributed in the Yellow Sea and Bohai Sea of China and the west coast of the Korean Peninsula. Different geographical populations represent potentially different genetic resources. To learn further the characteristics of different geographical population, crosses among two wild and three farmed populations were produced. The two wild populations were from the Yellow Sea and Bohai Sea （WYP）, and the west coast of the Korean Peninsula and coast （WKN）. The three farmed populations included the offspring of first generation of wild shrimp from coast in Korea （FKN）, the Huang Hai （the Yellow Sea in Chinese） No.1 （HH1）, and JK98. The phenotypes growth and survival rates of these populations were compared to confirm the feasibility for crossbreeding. The body length （BL）, carapace length （CL）. carapace width （CW）, height of the second and third abdominal segment （HST）, width of the second and third abdominal segment （WST）, length of the first abdominal segment （LF）, length of the last abdominal segment （LL）, live body weight （BW）, and survival rate were measured, Different combinations were statistically performed with ANOVA and Duncan＇s Multiple Range Test. The results show that the survival rate of JK98（♀）×WKN（♂） was the highest, followed by WYP（♀）×WKN（♂）, FKN（♀）×WYP（♂）, FKN（♀）×HHI（♂） and WYP（♀）×FKN（♂）; the body weight of FKN（♀）×HHI（♂） was the highest, followed by FKN（♀）×WYP（♂）, WYP（♀）×WKN（♂）, WYP（♀）×FKN（♂） and JK98（♀）×WKN（♂）; the total length had the same ranking as the body weight. All growth traits in hybrids JK98（♀）×WKN（♂） were the lowest among all combinations. F1 hybrids had significant difference （P〈0.05） in BL, CL, HST, LL, and BW; and insignificant difference （P〉0.05） in other growth traits and survival rate. The results of Duncan＇s Multiple Range Test are that BL and CL of J     

First record of <Emphasis Type="Italic">Paraonella platybranchia</Emphasis> (Hartman, 1961) (Polychaeta: Paraonidae) from the Yellow Sea

This paper reports for the first time a species of polychaetous family Paraonidae, Paraonella platybranchia (Hartman, 1961), a rare species found previously only in the coast near the boundary between the United States and Mexico, and it was recently discovered in the samples collected in 1982 from Chinese side of the Yellow Sea. This is the first record of the species in China as well as in the Northwest Pacific. Supported by the Knowledge Innovation Program of the Chinese Academy of Sciences (No. O72715)     

Variability of surface velocity in the Kuroshio Current and adjacent waters derived from Argos drifter buoys and satellite altimeter data

By combining Argos drifter buoys and TOPEX/POSEIDON altimeter data, the time series of sea-surface velocity fields in the Kuroshio Current (KC) and adjacent regions are established. And the variability of the KC from the Luzon Strait to the Tokara Strait is studied based on the velocity fields. The results show that the dominant variability period varies in different segments of the KC: The primary period near the Luzon Strait and to the east of Taiwan Island is the intra-seasonal time scale; the KC on the continental shelf of the ECS is the steadiest segment without obvious periodicity, while the Tokara Strait shows the period of seasonal variability. The diverse periods are caused by the Rossby waves propagating from the interior ocean, with adjustments in topography of island chain and local wind stress. Supported by the National Basic Research Program of China (973 Program, Nos. 2007CB411804, 2005CB422303), the NSFC (No. 40706006), the Key Project of International Science and Technology Cooperation Program of China (No. 2006DFB21250) and the “111 Project” (B07036), the Program for New Century Excellent Talents in University (NECT-07-0781)     

Screen and effect analysis of immunostimulants for sea cucumber, <Emphasis Type="Italic">Apostichopus japonicus</Emphasis>

Immunostimulants may improve disease resistance of aquaculture animals by promoting the nonspecific immunity response of the organisms. Five types of saccharides, including chitosan, yeast polysaccharide, burdock oligosaccharide, seaweed polysaccharide and lentinus edodes polysaccharide, were screened for potential use as immunostimulants by using spectrophotometry. The saccharides were injected into Apostichopus japonicus, a sea cucumber, and the lysozyme and superoxide dismutase (SOD) activities of the coelomic fluid and epidermal slime were monitored in six consecutive days. The results show that the lysozyme activity of the animal’s coelomic fluid was significantly stimulated on day 2, day 4 and day 6 after the injection of the saccharides (P<0.05). The effects of chitosan and yeast polysaccharide were the most notable. The lysozyme activity of the epidermal slime was significantly increased by chitosana, yeast polysaccharide, seaweed polysaccharide, and burdock oligosaccharide on day 1 and day 2 after the injection (P<0.05). The SOD activity of the coelomic fluid was significantly promoted by the saccharides on day 2 and day 4 post-injection (P<0.05), while the SOD activity of the epidermal slime increased on day 2. These findings indicate that chitosan and yeast polysaccharide are the most effective immunostimulants and potential healthy anti-disease feedstuff for A. japonicus. Supported by the National Key Technology Research and Development Program (863 Program, No. 2006BAD09A06) and the Special Fund of Chinese Central Government for Basic Scientific Research Operations in Commonweal Research Institutes (No. 02-2007B03)     

Effects of heavy metals （Pb^2＋ and Cd^2＋） on the ultrastructure, growth and pigment contents of the unicellular cyanobacterium Synechocystis sp. PCC 6803

The unicellular cyanobacterium Synechocystis sp. PCC 6803, a model organism known for its unique combination of highly desirable molecular genetic, physiological and morphological characteristics, was employed in the present study. The species was cultured in BG11 liquid medium contained various initial concentrations of Pb²⁺ and Cd²⁺ (0, 0.5, 1, 2, 4, 6 and 8 mg/L). The experiment was conducted for six days and the metal induced alterations in the ultrastructure, growth and pigment contents were assessed. Alterations in the ultrastructure of the Synechocystis sp. PCC 6803 cells became evident with the increased (>4 mg/L Pb²⁺) metal concentration. The photosynthetic apparatus (thylakoid membranes) were found to be the worst affected. Deteriorated or completely destroyed thylakoid membranes have made large empty spaces in the cell interior. In addition, at the highest concentration (8 mg/L Pb²⁺), the polyphosphate granules became more prominent both in size and number. Despite the initial slight stimulations (0.2, 3.8 and 6.5% respectively at 0.5, 1 and 2 mg/L Pb²⁺), both metals inhibited the growth in a dose-dependent manner as incubation progressed. Pigment contents (chlorophyll α, β carotene and phycocyanin) were also decreased with increasing metal concentration. Cells exposed to 6 mg/L Pb²⁺, resulted in 36.56, 37.39 and 29.34% reductions of chlorophyll α, β carotene and phycocyanin respectively over the control. Corresponding reductions for the same Cd²⁺concentrations were 57.83, 48.94 and 56.90%. Lethal concentration (96 h LC₅₀) values (3.47 mg/L Cd²⁺ and 12.11 mg/L Pb²⁺) indicated that Synechocystis sp. PCC 6803 is more vulnerable to Cd²⁺ than Pb²⁺. Supported by the Chinese Scholarship Council